Skip to main content

Advertisement

SpringerLink
Log in

The suitability of different DHB isomers as matrices for the MALDI-TOF MS analysis of phospholipids: which isomer for what purpose?

  • Article
  • Published:
European Biophysics Journal Aims and scope Submit manuscript

Abstract

Although the analysis of large biomolecules is the prime application of matrix-assisted laser desorption and ionization time-of-flight mass spectrometry (MALDI-TOF MS), there is also increasing interest in lipid analysis. Since lipids possess relatively small molecular weights, matrix signals should be as small as possible to avoid overlap with lipid peaks. Although 2,5-dihydroxybenzoic acid (DHB) is an established MALDI matrix, the question whether just this isomer is ideal for lipid analysis was not yet addressed. UV absorptions of all six DHB isomers were determined and their laser desorption spectra recorded. In addition, all isomers were used as matrices to record positive and negative ion mass spectra of selected phospholipids (phosphatidylcholine and -serine): In the order 2,5-, 2,6-, 2,3- and 2,4-DHB, the quality of the positive ion lipid spectra decreases. This correlates well with the decreasing acidity of the applied DHB isomers. The 3,4- and 3,5- isomers give only very weak positive ion signals especially of acidic lipids. In contrast, the most suitable matrices in the negative ion mode are 2,5-, 2,4- and 3,5-DHB. 2,6-DHB does not provide any signal in the negative ion mode due to its marked acidity. Finally, differences in the crystallization behavior of the pure matrix and the matrix/lipid co-crystals were also monitored by atomic force microscopy (AFM): 2,5-DHB gave the smallest crystals and the skinniest layer. It is concluded that basically all DHB isomers can be used as MALDI matrices but the 2,5-isomer represents the most versatile compound.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Al-Saad KA, Zabrouskov V, Siems WF, Knowles NR, Hannan RM, Hill HH Jr (2003) Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry of lipids: ionization and prompt fragmentation patterns. Rapid Commun Mass Spectrom 17:87–96

    Article  Google Scholar 

  • d’ Ans J, Lax E, Lechner MD (1992) Physikalisch-chemische Daten. Springer, Berlin Heidelberg New York

    Google Scholar 

  • Binnig G, Quate CF, Gerber C (1986) Atomic force microscope. Phys Rev Lett 56:930–933

    Article  ADS  Google Scholar 

  • Cristoni S, Bernardi LR (2003) Development of new methodologies for the mass spectrometry study of bioorganic macromolecules. Mass Spectrom Rev 22:369–406

    Article  Google Scholar 

  • Cvacka J, Svatos A (2003) Matrix-assisted laser desorption/ionization analysis of lipids and high molecular weight hydrocarbons with lithium 2,5-dihydroxybenzoate matrix. Rapid Commun Mass Spectrom 17:2203–2207

    Article  Google Scholar 

  • Estrada R, Yappert MC (2004) Alternative approaches for the detection of various phospholipid classes by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. J Mass Spectrom 39:412–422

    Article  Google Scholar 

  • Gut IG (2004) DNA analysis by MALDI-TOF mass spectrometry. Hum Mutat 23:437–441

    Article  MathSciNet  Google Scholar 

  • Hao C, Ma X, Fang S, Liu Z, Liu S, Song S, Liu J (1998) Positive and negative-ion matrix-assisted laser desorption/ionization mass spectrometry of saccharides. Rapid Commun Mass Spectrom 12:345–348

    Article  Google Scholar 

  • Harvey DJ (1995) Matrix-assisted laser desorption/ionization mass spectrometry of phospholipids. J Mass Spectrom 30:1333–1346

    Article  Google Scholar 

  • Harvey DJ (1999) Matrix-assisted laser desorption/ionization mass spectrometry of carbohydrates. Mass Spectrom Rev 18:349–450

    Article  Google Scholar 

  • Hillenkamp F, Karas M, Beavis RC, Chait BT (1993) Matrix-assisted laser desorption/ionization mass spectrometry of biopolymers. Anal Chem 63:1193A–1203A

    Google Scholar 

  • Horneffer V, Dreisewerd K, Lüdemann H-C, Hillenkamp F, Läge M, Strupat K (1999) Is the incorporation of analytes into matrix crystals a prerequisite for matrix-assisted laser desorption/ionization mass spectrometry? A study of five positional isomers of dihydroxybenzoic acid. Int J Mass Spectrom 185–187:859–870

    Google Scholar 

  • Horneffer V, Forsmann A, Strupat K, Hillenkamp F, Kubitscheck U (2001) Localization of analyte molecules in MALDI preparations by confocal laser scanning microscopy. Anal Chem 73:1016–1022

    Article  Google Scholar 

  • Karas M, Ehring H, Nordhoff E, Stahl B, Strupat K, Hillenkamp F, Grehl M, Krebs B (1993) Matrix-assisted laser desorption ionization mass spectrometry with additives to 2,5-dihydroxybenzoic acid. Org Mass Spectrom 28:1476–1481

    Article  Google Scholar 

  • Knochenmuss R, Zenobi R (2003) MALDI ionization: the role of in-plume processes. Chem Rev 103:441–452

    Article  Google Scholar 

  • Krause J, Stoeckli M, Schlunegger UP (1996) Studies on the selection of new matrices for ultraviolet matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Rapid Commun Mass Spectrom 10:1927–1933

    Article  Google Scholar 

  • Li YL, Gross ML, Hsu FF (2005) Ionic-liquid matrices for improved analysis of phospholipids by MALDI-TOF mass spectrometry. J Am Soc Mass Spectrom 16:679–682

    Article  Google Scholar 

  • Marto JA, White FM, Seldomridge S, Marshall AG (1995) Structural characterization of phospholipids by matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance mass spectrometry. Anal Chem 67:3979–3984

    Article  Google Scholar 

  • Müller M, Schiller J, Petković M, Oehrl W, Heinze R, Wetzker R, Arnold K, Arnhold J (2001) Limits for the detection of (poly-)phosphoinositides by matrix-assisted laser desorption and ionization time-of-flight mass spectrometry (MALDI-TOF MS). Chem Phys Lipids 110:151–164

    Article  Google Scholar 

  • Petković M, Schiller J, Müller M, Benard S, Reichl S, Arnold K, Arnhold J (2001) Detection of individual phospholipids in lipid mixtures by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry: phosphatidylcholine prevents the detection of further species. Anal Biochem 289:202–216

    Article  Google Scholar 

  • Raju NP, Mirza SP, Vairamani M, Ramulu AR, Pardhasaradhi M (2001) 5-Ethyl-2-mercaptothiazole as matrix for matrix-assisted laser desorption/ionization of a broad spectrum of analytes in positive and negative ion mode. Rapid Commun Mass Spectrom 15:1879–1884

    Article  Google Scholar 

  • Reinders J, Lewandrowski U, Moebius J, Wagner Y, Sickmann A (2004) Challenges in mass spectrometry-based proteomics. Proteomics 4:3686–3703

    Article  Google Scholar 

  • Schiller J, Arnhold J, Benard S, Müller M, Reichl S, Arnold K (1999) Lipid analysis by matrix-assisted laser desorption and ionization mass spectrometry: a methodological approach. Anal Biochem 267:46–56

    Article  Google Scholar 

  • Schiller J, Arnold K (2000a) Mass spectrometry in structural biology. In: Meyers RA (ed) Encyclopedia of analytical chemistry. John Wiley, Chichester pp 559–585

    Google Scholar 

  • Schiller J, Arnhold J, Glander HJ, Arnold K (2000b) Lipid analysis of human spermatozoa and seminal plasma by MALDI-TOF mass spectrometry and NMR spectroscopy—effects of freezing and thawing. Chem Phys Lipids 106:145–156

    Article  Google Scholar 

  • Schiller J, Zschörnig O, Petković M, Müller M, Arnhold J, Arnold K (2001) Lipid analysis of human HDL and LDL by MALDI-TOF mass spectrometry and 31P-NMR. J Lipid Res 42:1501–1508

    Google Scholar 

  • Schiller J, Süß R, Petković M, Zschörnig O, Arnold K (2002) Negative-ion matrix-assisted laser desorption and ionization time-of-flight mass spectra of complex phospholipid mixtures in the presence of phosphatidylcholine: a cautionary note on peak assignment. Anal Biochem 309:311–314

    Article  Google Scholar 

  • Schiller J, Süß R, Arnhold J, Fuchs B, Leßig J, Müller M, Petković M, Spalteholz H, Zschörnig O, Arnold K (2004) Matrix-assisted laser desorption and ionization time-of-flight (MALDI-TOF) mass spectrometry in lipid and phospholipid research. Prog Lipid Res 43:449–488

    Article  Google Scholar 

  • Schiller J, Süß R, Fuchs B, Müller M, Zschörnig O, Arnold K (2006) Recent applications of MALDI-TOF mass spectrometry and 31P NMR spectroscopy in lipid and phospholipid research. Future Lipidol 1:115–125

    Article  Google Scholar 

  • Tanaka T, Tsutsui H, Hirano K, Koike T, Tokumura A, Satouchi K (2004) Quantitative analysis of lysophosphatidic acid by time-of-flight mass spectrometry using a phosphate-capture molecule. J Lipid Res 45:2145–2150

    Article  Google Scholar 

  • Tsarbopoulos A, Karas M, Strupat K, Pramanik BN, Nagabhushan TL, Hillenkamp F (1994) Comparative mapping of recombinant proteins and glycoproteins by plasma desorption and matrix-assisted laser desorption/ionization mass spectrometry. Anal Chem 66:2062–2070

    Article  Google Scholar 

  • Wu KJ, Odom RW (1998) Characterizing synthetic polymers by MALDI MS. Anal Chem 70:456A–461A

    Google Scholar 

  • Yassin FH, Marynick DS (2005) Computational estimates of the gas-phase basicities, proton affinities and ionization potentials of the six isomers of dihydroxybenzoic acid. Mol Phys 103:183–189

    Article  Google Scholar 

Download references

Acknowledgments

This work was supported by the Deutsche Forschungsgemeinschaft (Schi 476/5-1) and the Federal Ministry of Education and Research (BMBF, Grant 982000-041).

Author information

Authors and Affiliations

  1. Institute of Medical Physics and Biophysics, Medical Faculty, University of Leipzig, Härtelstr. 16/18, 04107, Leipzig, Germany

    Jürgen Schiller, Rosmarie Süß, Beate Fuchs, Matthias Müller, Marijana Petković, Olaf Zschörnig & Hanka Waschipky

Authors
  1. Jürgen Schiller
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Rosmarie Süß
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Beate Fuchs
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Matthias Müller
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Marijana Petković
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Olaf Zschörnig
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Hanka Waschipky
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jürgen Schiller.

Additional information

Dedicated to Prof. Dr. Klaus Arnold on the occasion of his 65th birthday.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Schiller, J., Süß, R., Fuchs, B. et al. The suitability of different DHB isomers as matrices for the MALDI-TOF MS analysis of phospholipids: which isomer for what purpose?. Eur Biophys J 36, 517–527 (2007). https://doi.org/10.1007/s00249-006-0090-6

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00249-006-0090-6

Keywords

Search

Navigation